Among the primary obstacles to generating tissue engineered blood vessels (TEBV) is the inability of seeded cells to produce sufficient amounts of cross-linked elastin to achieve the mechanical characteristics of normal arteries. The objective of this proposal is to optimize conditions for adult smooth muscle cell proliferation, alignment and elastogenesis to achieve rapid development of TEBV in culture. A genetic engineering strategy based on overexpression of a splice variant of the proteoglycan versican, V3, induces elastogenesis in adult mammalian cells both in vitro and in vivo, but attenuates cell proliferation. The objectives of this proposal are to generate regulatable vectors for expression of V3 and other elastogenic genes so that the magnitude and timing of elastin synthesis can be controlled. We hypothesize that this system will enable optimization of cell coverage and elastin deposition on biomaterial scaffolds. To provide mechanical support and promote alignment, cells will be seeded onto grooved collagen membranes. Sheets of aligned cells will be formed into tubular constructs. The constructs will be subjected to radial strain in a bioreactor and the mechanical contribution of cell pre-alignment and elastogenesis will be assessed. [unreadable] [unreadable]